| AIMS Malignant lymphoma(ML), which is complicated in classification, bad in prognosis and extremely difficult in clinical diagnosis, is one of the top ten malignant tumors in our country and its misdiagnosis rate is as high as 30%. At the present time, the diagnosis of the ML is mainly depending on HE morphological study and immunological histochemistry and the ML diagnosis scheme, which is based on the HE morphological study and aided by immunological histochemistry and gene diagnosis, has been established in some famous hospitals. The final diagnosis rate is higher than 95% . However, the results of the immunological histochemistry often turn out to be false positive or false negative because the CD20A CD45RO Ab LCAAb are affected by various factors., need some more stable Ab; There are also many other problems, such as the reagents are not standardized, the conditions and methods are not unified, the discovery rates reported is inconsistent, the results are insecure, the gene diagnosis is hard to popularize and so on. Aiming at these problems mentioned above, we will make further researches on the following two aspects. First on immunological histochemistry, we draw an research on the express of B-cell-specific activator protein(BSAP) and CD20 during the lymphoproliterative disease and demonstrate the possibility of the BSAP as the clinical lymphoma identification antibody. Second on gene diagnosis, we draw an analogy among the multiple influences upon the HE dyeing and DNA preservation after the fixation of the fresh lymphoid tissue with different fixatives, such as 95% alcohol, 10% formaldehyde solution, neutral formaldehyde buffer, 4% citromint-O.lmol/L phosphoric acid buffer. Through this analogy we will seek the best fixation reagent and clarify its optimal fixing time andtemperature.METHODS (1) We observed the pathological morphology of 63lymphoproliferative diseases and classify them with the new WorldHealth Organization (WHO) classification, being integrated bywith CD45RO,. Then we maked an analogy of BSAP and CD20 with SABCmethods of immunological histochemistry; (2) We collected 30 casesof fresh tonsils tissue and after they were fixed (with 95% alcohol,10% formaldehyde solution, formaldehyde buffer, 4% formitrol etcrespectively and synchronously), paraffin-embeded, sliced and HEdying, we classified their tissue types and compare their dyingresults. 15 cases were fixed for different hours(1h 2h 4h81 24h 48h 72h) but in the same temperature (20癈) and the other15 cases were fixed for 8h in different temperatures (-20℃ ?℃ 20℃). We extracted DNA of these cases, tested the OD valueand maked an analogy of PCR gel electrophoresis.RESULTS (1) In the 63 cases of the lymphoproliferative disease,BSAP and CD20 are expressed on the surface of B lymphocytes of the9 cases of the lymph nodes reactive hyperplasia and the tumor cellsof the 34 cases of the B cell lymphoma. But the middle and strongpositive express rate of BSAP (69.8%) is higher than that of CD20(53. 5%) but there is no significant difference between the twogroups (P>0.05) . the express rate on the surface of the H/RS tumorcells of the 6 cases of Hodgkin lymphoma is 83.2%, and the BSAPis not expressed on the surface of the tumor cell of all the 14cases of the T cell lymphoma; (2)After fixing 30 cases of freshtonsils tissue, paraffin-embeded, sliced and HE dying, we findthat the 10% formaldehyde buffer and neutral morbicid bestpreserve the tissue structure and the dying color form a sharpcontrast. The tissue that was fixed with 95% alcohol is crisp andhard for slicing. The tissue structure under microscope is proneto transfigure. After the multiple study of the OD value and theamount of the DNA refinements, little difference of the density and ratio of the DNA was observed on the four fixed groups and the control groups which were fixed for different hours(1 IK 21u 4fK 8h, 24h, 48h, 72h) and in different temperatures(-20℃ , ?℃> 20癈) a...
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